栽培稻—药用野生稻杂种F1及回交后代的基因组原位杂交鉴定

以ｄｉｇｏｘｉｇｅｎｉｎ标记的药用野生稻总ＤＮＡ作探针,未标记的栽培稻总ＤＮＡ封阻,地栽培稻－药用野姓稻Ｆ１及回交皇代材料根尖体细胞染色体制片进行基因组原位杂交,鉴定出３个双单体异源附加系和４个单体异附加系。当标记探针用ａｎｔｉ－ｃｄｉｇｏｘｉｇｅｎｉｎ－ＰＯＤ和ＤＡＢ检测时,药用生稻染色体显棕色,栽培稻染色体则因Ｇｉｅｍｓａ.地染而显浅蓝色;标记探针用ａｎｔｉ－ｄｉｇｏｘｉｇｅｎｉｎ－ＦＩＴＣ检     

药用野生稻转育后代一个抗白叶枯病新基因的定位

从药用野生稻渗入后代选育的水稻株系B5表现为高抗褐飞虱、白背飞虱和白叶枯病。对B5与籼稻品种明恢63杂交组合的187个重组自交系(RILs)进行了抗白叶枯病接种鉴定,采用分离集团分析法(Bulked Segregant Analysis,BSA),在第1染色体上筛选到与水稻抗白叶枯病基因相连锁RFLP分子标记。利用RILs抗病性表现型鉴定资料和构建的分子标记连锁图谱,将抗白叶枯病基因定位在第1染色体短臂的C904和R596之间,这两个分子标记间遗传距离为1．3cM。该基因对RILs群体抗病性变异的贡献率为52．96％,是一效应值较大的主效基因。这一抗白叶枯病基因不同于已报道的抗白叶枯病基因的位点,因此将其命名为Xa29(t)。     

基因组原位杂交鉴定栽培稻与广西药用野生稻杂交后代染色体构成

广西药用野生稻（Ｏｒｙｚａ ｏｆｆｉｃｉｎａｌｉｓ）具有多种优良特性,是水稻遗传育种的重要种质资源之一。本实验以ｂｉｏｔｉｎ标记的的药用野生稻总ＤＮＡ作探针,未标记的载培稻（Ｏｒｙｚａｘａｔｉｖａ）总ＤＮＡ作封阻,以ＨＰ－ＤＡＢ系统进行信号检测,对栽培稻与广西药用野生稻的杂种Ｆ１植株的根尖染色体制片进行了基因组原位杂交。采用封阻比例１：２０～３０时,杂交效果较９为理想,药用野生稻的１２条染色体显深     

基因组原位杂交鉴定栽培稻与广西药用野生稻杂交后代染色体构成

广西药用野生稻 (Oryza officinalis)具有多种优良特性 ,是水稻遗传育种的重要种质资源之一。本实验以 Biotin标记的药用野生稻总 DNA作探针 ,未标记的栽培稻 (Oryzasativa)总 DNA作封阻 ,以 HRP- DAB系统进行信号检测 ,对栽培稻与广西药用野生稻的杂种F1植株的根尖染色体制片进行基因组原位杂交。采用封阻比例 1∶ 2 0～ 30时 ,杂交效果较为理想 ,药用野生稻的 1 2条染色体显深棕色 ,而栽培稻的 1 2条染色体着色很浅。在有丝分裂间期的细胞核中 ,也检测到大量杂交信号分布于核的周边。     

野生稻DNA导入水稻后代的SSR检测分析

以宁夏地区水稻主栽品种‘宁粳16号’、‘宁粳23号’为受体,普通野生稻(Oryza rufipogon,2n=2x=24, AA)为供体,利用花粉管通道导人法与茎注射法相结合的导入方法,将普通野生稻DNA导入宁夏水稻栽培品种, 对筛选出的21份形态、农艺性状典型变异的D2代材料及其对照利用SSR分子标记进行分子鉴定,结果表明,有 17份变异材料扩增出野生稻特有的DNA片段,而2个对照均不能扩增出野生稻特异的DNA片段,证明这些材料就是野生稻DNA片段导入的后代。另外有些材料出现受体DNA条带的减少,可能是由于野生稻DNA片段整合到水稻基因组中该引物结合位点或附近,原有的结合位点破坏而造成的。     

大白菜-结球甘蓝5号和8号单体异附加系的筛选和鉴定

为建立成套的大白菜-结球甘蓝异附加系,以大白菜-结球甘蓝异源三倍体（AAC,2n=3x=29）与二倍体大白菜（AA,2n=2x=20）回交世代BC1为材料,对其进行细胞学观察并从中筛选单体异附加系。结果表明,BC1群体植株发生了染色体数目变异,变异范围为2n=20～29,其中以2n=23～24的非整倍体植株居多,占总观察株数的51．44％;对2个2n=21的植株进行核型分析,初步鉴定为大白菜-结球甘蓝5号和8号单体异附加系,分别命名为CO-5—1和CO-8—2。     

珍稀濒危植物--云南药用野生稻自然生态群的新发现及其特性

野生稻包括云南药用野生稻(Oryza officinalis Wall.ex Watt.,染色体组CC,染色体数2n=24)是宝贵的基因资源库,自然分布急剧减少。2001年,在云南耿马孟定遮甸新发现了1个药用野生稻分布点,该地药用野生稻植株高度可塑性大,可以根据密度和荫蔽情况自动大幅度调节株高;比其它生态群的结实率高;而其自然发病和人工接种鉴定发现抗稻瘟病能力中等偏上,抗白叶枯病害能力强,抗螟虫和稻飞虱能力也很强。品质分析结果表明该居群药用野生稻蛋白质含量比其它野生稻的高,而直链淀粉含量偏低,说明品质较好。本发现对采取有效措施进行药用野生稻原生地保护,以及试验结果对药用野生稻有利基因的发掘和利用都有重要参考价值。     

云南药用野生稻核基因组细菌人工染色体(BAC)文库的构建与分析

通过云南药用野生稻核基因组BAC文库的构建,保存处于濒危状态的云南药用野生稻遗传资源,该文库包含27 500个克隆,随机挑取80个克隆检测,插入片段平均大小为80kb,文库容量相当于水稻基因大小的5.1倍。     

药用野生稻抗褐飞虱基因的RAPD标记研究

以药用野生稻1665(O.officinals)和栽培稻桂99远缘杂交的抗褐飞虱近等基因系B3F4分离群体为材料,利用随机引物对其抗褐飞虱基因进行了RAPD标记研究,获得3个与抗褐飞虱基因表现连锁的RAPD分子标记(S229703、S403783和S11591408)。采用MAPMAKER/EXP.Version 3.0分析,结果表明3个RAPD分子标记与抗褐飞虱基因属于同一连锁群,覆盖大小为18.1cm,标记间平均距离为4.53cm,为下一步水稻抗褐飞虱基因的精确定位和克隆奠定坚实的基础。     

单体异附加系花药培养创制小麦- 中间偃麦草纯合易位系

利用单体异附加系花药培养细胞工程途径,诱导小麦与中间偃麦草发生染色体易位,通过细胞学分析、荧光原位杂交(F ISH)和SSR鉴定出纯合易位系.研究结果表明,经单体异附加系花药培养创制出1个小麦-中间偃麦草纯合易位系99-803;其花粉母细胞(PM C s)减数分裂中期I染色体构型为18.42个环状二价体 2.57个棒状二价体 0.01个单价体;中间偃麦草的7A i-1染色体与小麦7A或7B染色体发生了非罗伯逊易位,且中间偃麦草易位片段较小;通过该途径获得纯合易位系的频率约为2%.以上结果表明,单体异附加系花药培养是一条向小麦转移异源染色体小片段(基因)的快速高效途径.     

Production and characterization of a complete set of individual chromosome additions from Oryza officinalis to Oryza sativa using RFLP and GISH analyses

Monosomic alien addition lines (MAALs) are valuable materials for comparative analyses of two distinct genomes, for elucidating introgression mechanisms, and for dissecting genes controlling complex traits. In the study reported here, MAALs of rice containing the complete genome of Oryza sativa and individual chromosomes of Oryza officinalis were produced. Interspecific hybridizations were made between O. sativa L. ssp. Japonica (CV, Hejiang 19, 2 n=24, AA) and O. officinalis (Acc. HY018, 2 n=24, CC). Two backcrosses were made to the cultivated rice to obtain BC₂F₁ plants. Through RFLP and GISH analyses, 25 MAALs (2 n=25, AA+1C) were identified and divided into 12 syntenic groups, designated MAALs 1–12. MAALs 1, 2, 3, 5, 7 and 10 were each represented by one plant, MAALs 8, 11 and 12 by two plants, MAALs 6 and 9 by four plants, and MAAL 4 by five plants. An ideogram of the C-genome of O. officinalis was constructed, based on GISH analysis of the interspecific hybrid and the MAALs. Comparative RFLP maps showed strong syntenic associations between the A-genomes and C-genomes. Chromosomal arrangements such as translocations and duplications were detected in different alien chromosomes of the MAALs. The complete set of O. officinalis MAALs generated here provides a novel manipulation platform for exploiting and utilizing the O. officinalis genome and carrying out genetic studies.     

Simple sequence repeat analyses of interspecific hybrids and MAALs of <Emphasis Type="Italic">Oryza </Emphasis><Emphasis Type="Italic">officinalis</Emphasis> and <Emphasis Type="Italic">Oryza sativa</Emphasis>

Wild rice is a valuable resource for the genetic improvement of cultivated rice (Oryza sativa L., AA genome). Molecular markers are important tools for monitoring gene introgression from wild rice into cultivated rice. In this study, Simple sequence repeat (SSR) markers were used to analyze interspecific hybrids of O. sativa-O. officinalis (CC genome), the backcrossing progenies and the parent plants. Results showed that most of the SSR primers (335 out of 396, 84.6%) developed in cultivated rice successfully amplified products from DNA samples of wild rice O. officinalis. The polymorphism ratio of SSR bands between O. sativa and O. officinalis was as high as 93.9%, indicating differences between the two species with respect to SSRs. When the SSR markers were applied in the interspecific hybrids, only a portion of SSR primers amplified O. officinalis-specific bands in the F(1) hybrid (52.5%), BC(1) (52.5%), and MAALs (37.0%); a number of the bands disappeared. Of the 124 SSR loci that detected officinalis-specific bands in MAAL plants, 96 (77.4%) showed synteny between the A and C-genomes, and 20 (16.1%) showed duplication in the C-genome. Sequencing analysis revealed that indels, substitution and duplication contribute to the diversity of SSR loci between the genomes of O. sativa and O. officinalis.     

Biochemical Analyses of the Antioxidative Activity and Chemical Ingredients in Eight Different Allium Alien Monosomic Addition Lines

We measured the antioxidant contents and antioxidative activities in eight Allium fistulosum–shallot monosomic addition lines (MAL; FF+1A–FF+8A). The high antioxidative activity lines (FF+2A and FF+6A) showed high polyphenol accumulation. These additional chromosomes (2A and 6A) would therefore have anonymous genes related to the upregulation of polyphenol production, the antioxidative activities consequently being increased in these MALs.     

Introgression of genes from Oryza officinalis Well ex Watt to cultivated rice,O. sativa L.

Summary Sterile AC hybrids between cultivated Oryza sativa (AA) and a distant wild species, O. officinalis (CC), were backcross to O. sativa. Most of the BC₁ progenies were allotriploid (AAC), a few were hypotriploid. AAC progenies were again backcrossed to O. sativa. BC₂ progenies consisting of disomic or aneuploid individuals were examined for the presence of O. officinalis traits. Eleven different traits from O. officinalis were identified in these progenies. Segregation data in the subsequent generations suggest that these traits are monogenic in nature. Two of these genes — for resistance to BPH and WBPH — are of value in rice improvement. The extremely low recovery of recombinant progenies is in agreement with the very low amount of pairing between A and C genomes. Because of this restricted recombination, the genotype of the recurrent parent was reconstituted after two backcrosses only. Thus, the BC₂ progenies look remarkably similar to O. sativa. Most of them are stable and fertile and also interfertile with other O. sativa breeding lines. Some of the BPH-and WBPH-resistant progenies are comparable in yield to the best O. sativa parents and are being evaluated as varietal possibilities.     

辽宁省近15年的部分水稻主栽品种的简单重复序列(SSR)多态性分析

应用简单重复序列(SSR)标记方法对辽宁省近15年的14个大面积种植的水稻品种进行遗传多样性分析的结果表明,17对引物共产生43个位点,其中多态性位点17个,平均每对SSR引物检测到2.53个,占位点总数的39.53%。用Nei’s公式计算水稻品种间的遗传距离,并以算术平均非加权聚类(UPGMA)法进行聚类分析并结合系谱分析结果表明,辽宁省近15年的水稻主栽品种遗传多样性不够丰富,多数品种间的亲缘关系较近,欲进一步提高产量还需拓宽遗传基础。     

水稻雄性不育恢复系明恢63的感光基因分析

水稻恢复系明恢63是中国应用面积最大、利用最广泛的恢复系。利用抽穗期感光性近等基因系EG0～EG7及ER～LR对明恢63进行的分析表明,明恢63在E1、E2、E3位点分别带有E1、e3、E3基因,在Se-1位点带无感光功能的Se-1^e基因。进一步用抽穗期QTL近等基因系NIL(Hd1)和NIL（Hd4)进行的研究表明,明恢63带有显性感光基因E1和无感光功能的Se-1^e基因,并推测明恢63带有能抑制E,基因表达的隐性抑制基因。认为籼型杂交稻抽穗期受不育系和恢复系感光基因及感光性抑制基因的共同作用。初步讨论了明恢63广适性的遗传基础。     

大白菜—结球甘蓝1号二体异附加系分子标记鉴定及其自交后代的性状分析

利用与抗霜霉病、抗芜菁花叶病毒基因连锁的SCAR标记和能够区分大白菜和结球甘蓝FLCs基因的特异引物对大白菜—结球甘蓝1号二体异附加系(AC1d)及其亲本进行分析,结果表明:AC1d及其双亲中均含有与抗霜霉病基因连锁的标记;AC1d和亲本结球甘蓝中含有与抗芜菁花叶病毒基因连锁的标记,亲本大白菜中无该标记;AC1d除具有4个大白菜Br FLCs基因外,同时还添加了结球甘蓝Bo FLC3基因。AC1d自交后代的株高、株展、球高、维生素C含量、可溶性蛋白质及其7种硫苷组分含量超出了亲本大白菜和结球甘蓝;叶形指数、可溶性糖含量和对小菜蛾的抗性等超出了亲本大白菜。为利用AC1d后代选育携有目标性状的易位系提供了依据。